LOCUS       sequence-148596-        7302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV packaging vector for Cre-dependent, hSyn driven KOR DREADD 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7302)
  AUTHORS   Vardy E, Robinson JE, Li C, Olsen RH, DiBerto JF, Giguere PM, 
            Sassano FM, Huang XP, Zhu H, Urban DJ, White KL, Rittiner JE, 
            Crowley NA, Pleil KE, Mazzone CM, Mosier PD, Song J, Kash TL, 
            Malanga CJ, Krashes MJ, Roth BL
  TITLE     A New DREADD Facilitates the Multiplexed Chemogenetic Interrogation 
            of Behavior.
  JOURNAL   Neuron. 2015 Apr 29. pii: S0896-6273(15)00289-5. doi: 
            10.1016/j.neuron.2015.03.065.
  PUBMED    25937170
REFERENCE   2  (bases 1 to 7302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2015 Apr 29. pii: S0896-6273(15)00289-5. doi: 
            10.1016/j.neuron.2015.03.065."
COMMENT     Sequence Label: pAAV-hSyn-dF-HA-KORD-IRES-mCitrine
FEATURES             Location/Qualifiers
     source          1..7302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(792..1502,1503..1505,1506..1508))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(834..855)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1183..1202
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1442..1463
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1505..1514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1515..2088
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1675..1694)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1904..1921
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             2602..2613
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(3247..3273)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3287..3320
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3371..3404
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3429..4017
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3482..3502)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3900..3911)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4049..4525
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4248..4267)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4565..4705
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4565..4694
                     /label=AAV2 ITR
     rep_origin      4780..5235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4867..4886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5077..5098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5252..5271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5371..5393
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5431..5449)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5517..5621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5622..5690,5691..6482)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5840..5859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6653..7241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7142..7161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccctatc acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
      841 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagttt ggactggtag
      901 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
      961 tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc
     1021 ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac
     1081 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     1141 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     1201 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
     1261 tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcaca tcaggccgta gccgaaggtg
     1321 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc
     1381 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     1441 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     1501 ctcaccatgg tggcttatca tcgtgttttt caaaggaaaa ccacgtcccc gtggttcggg
     1561 gggcctagac gtttttttaa cctcgactaa acacatgtaa agcatgtgca ccgaggcccc
     1621 agatcagatc ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg
     1681 cttgaggaga gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt
     1741 tgtgccgcct ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg
     1801 ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa
     1861 gcttccagag gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga
     1921 ggggaaagac ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca
     1981 ttgccaaaag acggcaatat ggtggaaaat aacatataga caaacgcaca ccggccttat
     2041 tccaagcggc ttcggccagt aacgttaggg gggggggagg gagaggggcg atatcgcggc
     2101 cgctcatact ggtttattca tcccatcgat gtccctcagg taagcaggat cctgaactgt
     2161 atttcggact ctgctagtgc tctgccgctc catcctcatc ttcagtggaa agcagaagtc
     2221 ccggaaacac cgcttgaagt tttcatcaag aaaggcgtag agaatgggat tcaggctact
     2281 gttggtatag cctaaggcga tgcagaagta atagctggag agagcagctg tgctgtggga
     2341 ggtgctcccc agagcctcca ccaggatgaa tatgtgaatg ggagtccagc agacgacgaa
     2401 gactgccacc accaccagga ccagtctggt gatcctacgc aggttgcgat ctttctctcg
     2461 ggagccagaa aggagccgga cgctcttgag acgcaggatc atcagggtgt agcagacgat
     2521 gatgatgagg acagggatca cgaaggcaaa gatgaagacg cagatcttca tgaagaggtc
     2581 ccaccaggag tagtcatcat ctgggaactg caaggagcac tcaatgacat cgacgtcttc
     2641 cctgactttg gtgcctccaa ggactattgc agagatgcca acagatgacg acagcagcca
     2701 gatgcagata ttgatgatct ttgccttcaa gggtgtgcgg aagtccaaag ccttcacggg
     2761 gtggcacacg gcaatgtagc ggtccacgct catcatggtc aaggtgaaga tgctggtgaa
     2821 catgttgtag taattaatgg aaattactat cttgcacagc acatccccaa aaggccagga
     2881 attcatcaag tagaccgtac tctgaaaggg catggttgta gtaactaaag catctgccaa
     2941 agccaggtta aatatgtaaa tgttggttgc tgtcttcatc tttgtgtatc ggatgatcac
     3001 gaacatgacc agcgagttgc ccaccaagcc cacgacgaac actacggagt agaccgccgt
     3061 gatgatgacc gggatggccg gggagatgtg cgcgggctcc agctgcgcgt cctccgagcc
     3121 ggcgctgccg ttgctgtcgg gctcggccca gccgggaaac caggcgctgc tgttgggggg
     3181 caggcaggcg ctcggggcgc aggtagggcc cggctccccg cggaagatct gaatcgggga
     3241 ttccatagcg taatctggaa catcgtatgg gtacatggtg gctagcataa cttcgtataa
     3301 agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat
     3361 ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc
     3421 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3481 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3541 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3601 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3661 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3721 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3781 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3841 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3901 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3961 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     4021 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     4081 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     4141 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     4201 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     4261 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4321 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4381 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4441 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4501 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4561 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4621 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4681 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4741 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4801 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4861 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4921 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4981 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5041 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5101 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     5161 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5221 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5281 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5341 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5401 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5461 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5581 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5641 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5701 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5761 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5821 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5881 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5941 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6001 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6061 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6121 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6181 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6241 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6301 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6361 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6421 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6481 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6541 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6601 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6781 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7021 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7201 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7261 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
